Rajasekaran Rohith, Galateo Thomas M, Xu Zhejing, Bolshakov Dennis T, Weix Elliott W Z, Coyle Scott M
Department of Biochemistry, University of Wisconsin-Madison, Madison, Wisconsin 53706, USA.
Integrated Program in Biochemistry Graduate Program, University of Wisconsin-Madison, Madison, Wisconsin 53706, USA.
bioRxiv. 2025 Feb 28:2025.02.28.640587. doi: 10.1101/2025.02.28.640587.
Radios and cellphones use frequency modulation (FM) of an oscillating carrier signal to reliably transmit multiplexed data while rejecting noise. Here, we establish a biochemical analogue of this paradigm using genetically encoded protein oscillators (GEOs) as carrier signals in circuits that enable continuous, real-time FM streaming of single-cell data. GEOs are constructed from evolutionarily diverse MinDE-family ATPase and activator modules that generate fast synthetic protein oscillations when co-expressed in human cells. These oscillations serve as a single-cell carrier signal, with frequency and amplitude controlled by GEO component levels and activity. We systematically characterize 169 ATPase/activator GEO pairs and engineer composite GEOs with multiple competing activators to develop a comprehensive platform for waveform programming. Using these principles, we design circuits that modulate GEO frequency in response to cellular activity and decode their responses using a calibrated machine-learning model to demonstrate sensitive, real-time FM streaming of transcription and proteasomal degradation dynamics in single cells. GEOs establish a dynamically controllable biochemical carrier signal, unlocking noise-resistant FM data-encoding paradigms that open new avenues for dynamic single-cell analysis.
收音机和手机利用振荡载波信号的频率调制(FM)来可靠地传输多路复用数据,同时抑制噪声。在此,我们建立了这种范式的生化模拟,在能够实现单细胞数据连续、实时FM流传输的电路中,使用基因编码的蛋白质振荡器(GEO)作为载波信号。GEO由进化上不同的MinDE家族ATP酶和激活剂模块构建而成,当在人类细胞中共表达时,这些模块会产生快速的合成蛋白质振荡。这些振荡作为单细胞载波信号,其频率和幅度由GEO组件水平和活性控制。我们系统地表征了169个ATP酶/激活剂GEO对,并设计了具有多个竞争性激活剂的复合GEO,以开发一个用于波形编程的综合平台。利用这些原理,我们设计了响应细胞活性来调制GEO频率的电路,并使用校准的机器学习模型对其响应进行解码,以展示单细胞中转录和蛋白酶体降解动力学的灵敏、实时FM流传输。GEO建立了一个动态可控的生化载波信号,开启了抗噪声FM数据编码范式,为动态单细胞分析开辟了新途径。